Jan Kreft wrote
I believe biofilm research in the future should be more quantitative.
Quantitative analysis of biofilm structure; beyond beautiful pictures!
Quantitative work on pheromones in biofilms.
That's my $.02.
Jan Kreft
Quantification beyond beautiful pictures is certainly one of the major
research goals in our laboratory. I think that there are three different
overall parameters that have to be considered when we talk about
quantification of biofilms. These are the organism (who is there), the
structure (where are they) and the interactions (what are the organisms
doing).
To quantify the fraction of different species, and to identify their
location in the biofilm is probably where we are right now. By the use of
fluorescent in situ hybridization techniques (FISH) is it possible to
identify specific species, and to determine their specific location in the
biofilm (showing that produces beautiful pictures).
However, I think that one of the real challenges comes when we want to
study the interactions occurring between different organisms and between
organisms and their local environments. One way to do that is by fusing
reporter genes (such as for example the gfp gene) to specific promoters
that respond to the environmental signals. At the moment this method allows
us to distinguish between low or high level of gene expression in the
individual cells, but to obtain quantitative information about the exact
expression level in each individual cell within the biofilm is more
complicated. To do that, problems with for example photo bleaching and the
fact that light scattering is influenced by the cell densities and EPS
concentrations, must be taken into account. We also need to develop image
analysis programs, that allows us to automatically detect individual cells
located even in tight bacterial clusters and, which furthermore allow us to
quantify the "exact" signal intensity emitted from each individual cell in
the biofilm.
So, much in the line of Poul Stoodleys comments I think that it is
important to join the different disciplines such as the mathematicians (to
develop image treatment programs), the physicists (to solve some of the
problems with photo bleaching etc.), and us, the microbial ecologists, who
want to understand the nature of these biofilms.
Bjarke
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* Bjarke Bak Christensen, PhD *
* Department of Microbiology Technical University of Denmark *
* Building 301 DK 2800 Lyngby Denmark *
* Phone: (+45) 45 25 67 27 FAX: (+45) 45 93 28 09 *
* E-mail: bbc at im.dtu.dk *
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